Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells.
Characterisation of K+ channels in human fetoplacental vascular smooth muscle cells.
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DOI：
10.1371/journal.pone.0057451
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Greenwood SL
中科院分区：
文献类型：
--
作者：
Brereton MF;Wareing M;Jones RL;Greenwood SL
充足的血流量通过胎盘绒毛膜板阻力动脉（CPAs）是氧气和营养物质转移到胎儿和成功妊娠所必需的。在非胎盘血管平滑肌细胞（SMCs）中，K+通道调节收缩、血管张力和血流。先前的研究表明，K+通道调节剂改变CPA音调，但没有区分内皮细胞和SMCs对K+通道的影响。在本研究中，我们制备了新鲜分离的正常妊娠CPASMCs，并研究了K+通道的表达和功能。采用酶解法从正常人足月胎盘中分离出cpasmc。纯度和表型经免疫细胞化学证实。使用全细胞膜片钳评估K+通道电流，并通过RT-PCR和免疫染色检测完整cpa和分离SMCs的mRNA和蛋白表达。分离的SMCs表达α-肌动蛋白，但不表达内皮细胞标志物CD31。cpasmc和完整CPAs表达h-caldesmon和非肌球蛋白重链-2；分别为收缩型和合成型SMCs的表型标记。全细胞电流被4-AP、TEA、肉毒毒素和iberiotoxin所抑制，这些毒素与功能Kv和BKCa通道有关。1-EBIO增强了全细胞电流，这些电流分别被TRAM-34消除，被apamin降低，表明IKCa和SKCa被激活。BKCa、IKCa和SKCa3 mRNA和/或蛋白在cpasmc和完整的cpa中表达。这项研究首次提供了在cpasmc中Kv、BKCa、IKCa和SKCa通道功能的直接证据。这些细胞表现出混合表型，暗示CPASMCs在控制胎儿胎盘血管阻力和血管发生方面具有双重作用。
Adequate blood flow through placental chorionic plate resistance arteries (CPAs) is necessary for oxygen and nutrient transfer to the fetus and a successful pregnancy. In non-placental vascular smooth muscle cells (SMCs), K+ channels regulate contraction, vascular tone and blood flow. Previous studies showed that K+ channel modulators alter CPA tone, but did not distinguish between effects on K+ channels in endothelial cells and SMCs. In this study, we developed a preparation of freshly isolated CPASMCs of normal pregnancy and investigated K+ channel expression and function. CPASMCs were isolated from normal human term placentas using enzymatic digestion. Purity and phenotype was confirmed with immunocytochemistry. Whole-cell patch clamp was used to assess K+ channel currents, and mRNA and protein expression was determined in intact CPAs and isolated SMCs with RT-PCR and immunostaining. Isolated SMCs expressed α-actin but not CD31, a marker of endothelial cells. CPASMCs and intact CPAs expressed h-caldesmon and non-muscle myosin heavy chain-2; phenotypic markers of contractile and synthetic SMCs respectively. Whole-cell currents were inhibited by 4-AP, TEA, charybdotoxin and iberiotoxin implicating functional Kv and BKCa channels. 1-EBIO enhanced whole cell currents which were abolished by TRAM-34 and reduced by apamin indicating activation of IKCa and SKCa respectively. BKCa, IKCa and SKCa3 mRNA and/or protein were expressed in CPASMCs and intact CPAs. This study provides the first direct evidence for functional Kv, BKCa, IKCa and SKCa channels in CPASMCs. These cells display a mixed phenotype implicating a dual role for CPASMCs in controlling both fetoplacental vascular resistance and vasculogenesis.
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影响因子：
20.1
作者：
Gollasch, M;Ried, C;Haller, H
通讯作者：
Haller, H
影响因子：
8.3
作者：
McNeish, AJ;Sandow, SL;Garland, CJ
通讯作者：
Garland, CJ
DOI：
10.1016/s0002-9378(11)80047-5
发表时间：
1991-02-01
影响因子：
9.8
作者：
MYATT, L;BREWER, A;BROCKMAN, DE
通讯作者：
BROCKMAN, DE
DOI：
10.1152/ajpheart.01033.2001
发表时间：
2002-12-01
影响因子：
4.8
作者：
Hampl, V;Bíbová, J;Archer, SL
通讯作者：
Archer, SL
影响因子：
7.3
作者：
Halliday, FC;Aaronson, PI;Gurney, AM
通讯作者：
Gurney, AM